| 预期应用 ELISA法定量测定人血清、血浆、细胞培养物上清或其它相关液体中白细胞介素23(IL-23)含量。 实验原理 本试剂盒应用双抗体夹心酶标免疫分析法测定标本中IL-23水平。用纯化的抗体包被微孔板,制成固相抗体,往包被单抗的微孔中依次加入IL-23、生物素化的抗人IL-23抗体、HRP标记的亲和素,经过彻底洗涤后用底物TMB显色。TMB在过氧化物酶的催化下转化成蓝色,并在酸的作用下转化成最终的黄色。颜色的深浅和样品中的IL-23呈正相关。用酶标仪在450nm波长下测定吸光度(OD值),计算样品浓度。  Intended use This immunoassay kit allows for the specific measurement of Human IL-23 concentrations in cell culture supernates, serum, and plasma. Introduction Interleukin-23 (IL-23) is a heterodimeric cytokine consisting of two subunits, one called p40, which is shared with another cytokine, IL-12, and another called p19 (the IL-23 alpha subunit). IL-23 is an important part of the inflammatory response against infection. It promotes upregulation of the matrix metalloprotease MMP9, increases angiogenesis and reduces CD8+ T-cell infiltration. Recently, IL-23 has been implicated in the development of cancerous tumors. In conjunction with IL-6 and TGF-β1, IL-23 stimulates naive CD4+ T cells to differentiate into a novel subset of cells called Th17 cells, which are distinct from the classical Th1 and Th2 cells. Th17 cells produce IL-17, a proinflammatory cytokine that enhances T cell priming and stimulates the production of proinflammatory molecules such as IL-1, IL-6, TNF-alpha, NOS-2, and chemokines resulting in inflammation. Knockout mice deficient in either p40 or p19, or in either subunit of the IL-23 receptor (IL-23R and IL12R-β1) develop less severe symptoms of multiple sclerosis and inflammatory bowel disease highlighting the importance of IL-23 in the inflammatory pathway. Interleukin 23 (IL-23) is a heterodimeric cytokine composed of two disulfide-linked subunits, a p19 subunit that is unique to IL-23, and a p40 subunit that is shared with IL-12. The p19 subunit has homology to the p35 subunit of IL-12, as well as to other single chain cytokines such as IL-6 and IL-11. The p40 subunit is homologous to the extracellular domains of the hematopoietic cytokine receptors. Human p19 cDNA encodes a 189 amino acid residue (aa) precursor protein with a putative 19 aa signal peptide and 170 aa mature protein. Human and mouse p19 share 70% aa sequence identity. Although p19 is expressed by activated macrophages, dendritic cells, T cells, and endothelial cells, only activated macrophages and dendritic cells express p40 concurrently to produce IL-23. The functional IL-23 receptor complex consists of two receptor Test principle This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IL-23 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-23 present is bound by the immobilized antibody. An enzyme-linked polyclonal antibody specific for IL-23 is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of IL-23 bound in the initial step. The color development is stopped and the intensity of the color is measured. |